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Thalassemia

Alpha ProMix Test Kit

Novel Alpha Thalassemia Test can be used for the detection of Alpha Thalassemia subtypes by addition of DNA sample and processing via thermal cycler using conventional PCR technique. Test reagent kit based on conventional PCR.

Easy-to-use test kit due to the low effort required to prepare the multiplex PCR master mix. Each component of the test kit is already present in the appropriate quantity. This leads to a higher rate of successful experiments.

THE DISEASE DEFINITION

α-Thalassaemia often occurs in tropical and subtropical malaria endemic areas. It is characterised by a reduced or completely absent synthesis of a-globin chains. This leads to the excess of g- or b-chains, which form the homotetramers Hb Barts’s (g4) and HbH (b4), respectively. Both Hb Bart’s and HbH bind O2 with high affinity and no cooperative effect, so they do not function as O2 transporters.
In addition, their solubility in erythroid cells is lower than that of HbA. These excess haemoglobins thus lead to erythrocyte membrane damage and haemolytic anaemia.
The a-globin chains are encoded by two, tightly coupled and almost identical genes. Thus, the normal complement of a-globin genes of the diploid chromosome set consists of four genes.
The clinical picture depends on the number of a-globin genes that are still functional. In contrast to b-thalassaemia, a-thalassaemia mutations are mostly deletions.
The spectrum includes intrauterine fetal death when all 4 a-globing genes are inactivated (Hb-Bart’s hydrops fetalis), thalassaemia intermedia when 3 genes are inactivated (HbH disease), thalassaemia minor when 2 are absent, and a clinically and haematologically elusive form when one a-globing gene is absent.

Alpha Promix Test Kit

Is a testing reagent designed based on the principle of Conventional PCR.
This Test Kit is used for the detection of alpha-thal gene abnormality, which is a cause and an inherited factor of thalassemia.
This Test Kit comprises all PCR reagents including Taq DNA Polymerase enzyme, dNTPs, Primer, MgCl2 and buffer in a form of ready-to-use PCR mastermix solution contained in PCR tube strip.
Can be used for the detection by adding DNA sample and enter the process of Thermal cycler using Conventional PCR technique.

HIGHLIGHTS

The Test Kit allows the user to perform detection more easily and also reduces the procedures in preparing multiplex PCR mastermix which is more complicated than general singleplex PCR.
The Test Kit allows the user to perform detection more easily and also reduces the procedures in preparing multiplex PCR mastermix which is more complicated than general singleplex PCR.

Testing Procedures

Analysis using gel electrophoresis.

Enter the process of PCR ampflification by thermal cycler (Conventional PCR).

Pipette DNA sample 1 ul (concentration 100-200 ng/ ul) Into Alpha Promix reagent strip.

Features of the Test Kit

Handy and easy to use Test Kit due to the less procedures In preparing multiplex PCR mastermix. Each component in the Test Kit is already present in appropriate amount. This resulted in higher rate of successful experiment.
The speed of the Test Kit experiment is high. This reduces the time for each experiment and make the results to be obtained much faster within a day time.
Reduce error rate and contamination that could occur while preparing PCR mastermix reaction.

Features and benefits of the Test Kit

Compositions of Alpha Promix Test Kit

Alpha Promix reagents are contained in PCR tube strip
3 test applications are provided including.

3 deletions Test

5 deletions alpha – Thai 1&2

Alpha variants Test

Details of Alpha Promix Test Kit

The Test Kit is divided into 3 applications to provide the user with options to select the suitable test as desired, which includes:

Alpha Promix

Note;
-NC = Negative control resulted in the appearance of only one internal control (beta actin) band (123 bp).

-PC mix = Positive control, the mixture of DNA fargment clone of each positive control deletion (not provided).

Alpha Promix

Note;
Control band 2222 bp (Top band) can be inierpreted a-deletion as heterozygous or homozygous

Alpha Promix

Note;
Band 961 bp (Top band) caused by the binding of the primers in this reaction is not specific (non-specific band), which cannot be interpreted to be heterozygous or homozygous. To report heterozygous or homozygous, the normal primer for that mutation must be further tested (this test kit cannot be classified).

Storage conditions – Highlights

Store at -20 to 4 degrees Celsius for up to 6 months.
Multiple freeze / thaws are not recommended.